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基于转录组数据的桔小实蝇微卫星位点信息分析

魏丹丹,石俊霞,张夏瑄,陈世春,魏冬,王进军**   

  1. (西南大学植物保护学院昆虫学及害虫控制工程重庆市重点实验室, 重庆 400716)
  • 出版日期:2014-06-18 发布日期:2014-06-18

Analysis of microsatellite loci from Bactrocera dorsalis based on transcriptome dataset.

WEI Dan-dan, SHI Jun-xia, ZHANG Xia-xuan, CHEN Shi-chun, WEI Dong, WANG Jin-jun   

  1. (Chongqing Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, China)
  • Online:2014-06-18 Published:2014-06-18

摘要:

以桔小实蝇为材料,从其转录组数据库中筛选功能微卫星(EST-SSR)序列,并进行SSR位点的信息分析.共获得1890个EST-SSR位点,可用于引物设计的SSR数为1296个.EST-SSR平均分布频率为1/10.21 kb,但这种分布频率在不同重复类型SSR之间相差很大.其中,三碱基重复SSR在该种昆虫的EST-SSR中出现的频率最高,结合其他文献推断三碱基重复可能是所有昆虫EST-SSR的优势类型.本文共设计了42对桔小实蝇EST-SSR引物,其中有18对引物可以扩增得到预期大小的条带.最后,探讨了基于转录组数据发掘昆虫SSR的前景和挑战,以及进行转录组EST-SSR筛选时应注意的问题.

 

Abstract:

The transcriptome database of the oriental fruit fly, Bactrocera dorsalis (Hendel), was used to identify the functional gene-microsatellite (EST-SSR) markers and to analyze the SSR loci information. In total, 1890 EST-SSR loci were identified, of which, 1296 SSR sequences could be used for primer design. The average distribution frequency of the transcriptomic SSRs was 1/10.21 kb. However, these distribution frequencies varied considerably among different types of repeat SSRs. The trinucleotide repeat SSRs were found to have the highest frequency among the different types of repeat SSRs in the EST-SSR of B. dorsalis. Combining with other literatures, we inferred that the trinucleotide repeat SSRs were the most abundant EST-SSR in all of insects. In this study, 42 pairs of EST-SSR primers were designed and 18 pairs produced amplification bands of expected sizes. According to the results of other related literatures, the practices and challenges of strategy for SSR isolation from insect transcriptome databases were discussed, and the problems which should be considered in the screening of insect transcriptomic EST-SSR were put forward.